The comprehensive mission governing the strategy of the Developmental Resource for Biophysical Imaging and Opto-Electronics (DRBIO) is the creation and facilitation of instrumentation technologies that advance capabilities for visualization and measurement of dynamic biomolecular and cellular processes. Thus, the core research aims to create innovations that are developed by "Biomedical Engineering" into "Technologies for the Study of Molecular and Cellular Structure and Function." It is the challenges of "impossible"- but crucial - biological problems that motivate each new direction of our core research, leading to developments of instrumentation to facilitate our collaborations and services, the dissemination of the resulting innovations, and to training of participants and clients to support the transfer of the new technologies to the biomedical community. This combination of instrument development and collaboration is efficient and synergistic, with frequent reciprocal intellectual exchanges between collaborators and staff creating new concepts and understanding for both groups. The program comprises three classes of projects: (1) Biomedical instrumentation engineering to advance the effectiveness of biomedical application of the technologies of multiphoton microscopy (MPM) and fluorescence correlation spectroscopy (FCS), invented and already featured in DRBIO. (2) New directions of development of photophysical technologies, focusing particularly on robust optical markers for sensitive single molecule observations, and on in vivo imaging of the intrinsic fluorescence of tissue components. (3) Venturesome biotechnology developments based on new technologies, including optical nanostructures, nonlinear photochemistry and correlation spectroscopy to address several crucial but refractory challenges of biomedical research.